ways to get around self activation in yeast 2 hybrid...

Frederik Boernke boernke at nospam.ipk-gatersleben.de
Mon Jun 29 13:40:45 EST 1998


Sai Iyer wrote:
> 
> hail all,
> 
>         my construct is a strong self activator when i use it as a BD fusion to
> fish interacting proteins from a cDNA library.  use of 3 amino triazole
> is not a option since the activation is immediate (in context to 2
> hybrid activation times) i.e in about 5 min.  so is there any other
> possible option that i can employ to fish out interacting proteins from
> a library using 2 hybrid?? i should mention that i dont have
> any candidate clones yet; so switching BDs and ADs is also not an option
> (yet)...thanks
> 
> --
> Sai Iyer (siyer at bs.jhmi.edu)
> Pre-doctoral fellow i.e lowly graduate student
> Dept. of Biological Chemistry
> Johns Hopkins University School of Medicine

Hi,

the only thing I can think of is making deletion constructs in order
to get rid of the selfactivating domain. However, this can also change
interacting properties.TMO the best way to solve your problem is to
switch to another method.

Good luck
Ricky

******************************************************************

Frederik Boernke
Research Group of  Molecular Plant Physiology
Institute for Plant Genetics and Crop Plant Research (IPK)
Corrensstr. 3
06466 Gatersleben
Tel.  039482 -5 321
Fax. 039482 -5 515
http://www.ipk-gatersleben.de



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